Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
NH4Cl | 1.0 | g | 0.978 | 18.293 | |
MgCl2 x 6 H2O | 1.0 | g | 0.978 | 4.813 | |
CaCl2 x 2 H2O | 0.4 | g | 0.391 | 2.662 | |
KH2PO4 | 0.4 | g | 0.391 | 2.876 | |
Wolfe's mineral elixir | 1.0 | ml | - | - | |
Yeast extract | 2.0 | g | 1.957 | - | |
Trypticase peptone
(BD BBL) |
2.0 | g | 1.957 | - | |
Sodium resazurin
(0.1% w/v) |
0.5 | ml | 4.9e-4 | 0.002 | |
Na2CO3 | 1.0 | g | 0.978 | 9.232 | |
Na-crotonate solution (1 M) | 20.0 | ml | - | - | |
2-Mercaptoethanesulfonic acid
(coenzyme M) |
0.5 | g | 0.489 | 3.44 | |
Wolin's vitamin solution (10x) | 1.0 | ml | - | - | |
Na2S x 9 H2O | 0.3 | g | 0.294 | 1.222 | |
Distilled water | 1000.0 | ml | - | - | |
1 Dissolve ingredients (except carbonate, crotonate, coenzyme M, vitamins and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into Hungate-type tubes or serum vials (e.g., 5 ml medium in Balch-type tubes) and autoclave. Add crotonate, coenzyme M, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of crotonate, coenzyme M and vitamins should be sterilized by filtration. Adjust pH of the complete medium to 7.0 - 7.2. | |||||
2 Note: Use 10% (v/v) as inoculum. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
MgSO4 x 7 H2O | 30.00 | g | 30 | 121.716 | |
MnSO4 x H2O | 5.00 | g | 5 | 29.583 | |
NaCl | 10.00 | g | 10 | 171.116 | |
FeSO4 x 7 H2O | 1.00 | g | 1 | 3.597 | |
CoCl2 x 6 H2O | 1.80 | g | 1.8 | 13.863 | |
CaCl2 x 2 H2O | 1.00 | g | 1 | 6.802 | |
ZnSO4 x 7 H2O | 1.80 | g | 1.8 | 6.26 | |
CuSO4 x 5 H2O | 0.10 | g | 0.1 | 0.401 | |
AlK(SO4)2 x 12 H2O | 0.18 | g | 0.18 | 0.379 | |
H3BO3 | 0.10 | g | 0.1 | 1.617 | |
Na2MoO4 x 2 H2O | 0.10 | g | 0.1 | 0.413 | |
(NH4)2Ni(SO4)2 x 6 H2O | 2.80 | g | 2.8 | 7.089 | |
Na2WO4 x 2 H2O | 0.10 | g | 0.1 | 0.303 | |
Na2SeO4 | 0.10 | g | 0.1 | 0.529 | |
Distilled water | 1000.00 | ml | - | - | |
1 First adjust pH to 1.0 with diluted H2SO4, then add and dissolve the salts. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Crotonic acid
(ALDRICH 113018) |
86 | g | 86 | 998.955 | |
NaOH
(10 N) |
100 | ml | - | - | |
Distilled water | 900 | ml | - | - | |
1 Dissolve crotonic acid in 800 ml distilled water, add around 100 ml of 10 N NaOH and adjust pH to around 7. Then add water to reach a volume of 1000 ml. Sterilize by filtration under 100% N2 gas atmosphere. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Biotin | 20 | mg | 0.02 | 0.082 |
Folic acid | 20 | mg | 0.02 | 0.045 |
Pyridoxine hydrochloride | 100 | mg | 0.1 | 0.486 |
Thiamine HCl | 50 | mg | 0.05 | 0.148 |
Riboflavin | 50 | mg | 0.05 | 0.133 |
Nicotinic acid | 50 | mg | 0.05 | 0.406 |
Calcium D-(+)-pantothenate | 50 | mg | 0.05 | 0.21 |
Vitamin B12 | 1 | mg | 1.0e-3 | 7.4e-4 |
p-Aminobenzoic acid | 50 | mg | 0.05 | 0.365 |
(DL)-alpha-Lipoic acid | 50 | mg | 0.05 | 0.242 |
Distilled water | 1000 | ml | - | - |
Last modified: | 31.08.23 |
Source: | DSMZ |
Medium type: | Complex medium |
Final pH: | 7.0 - 7.2 |
Equipment needed: | Autoclave, Filter, Gassing station, Hungate tubes |
Edit in Medium builder |
Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .
Compound | Conc. [g/L] | Conc. [mM] |
---|---|---|
Yeast extract | 1.957 | - |
Trypticase peptone | 1.957 | - |
Crotonic acid | 1.683 | 19.549 |
NH4Cl | 0.978 | 18.293 |
Na2CO3 | 0.978 | 9.232 |
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